An alternate splicing, variant of the human telomerase catalytic subunit inhibits telomerase activity

X. Yi, D. M. White, D. L. Aisner, J. A. Baur, W. E. Wright, J. W. Shay

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Telomerase, a cellular reverse transcriptase, adds telomeric repeats to chromosome ends. In normal human somatic cells, telomerase is repressed and telomeres progressively shorten, leading to proliferative senescence. Introduction of the telomerase (hTERT) cDNA is sufficient to produce telomerase activity and immortalize normal human cells, suggesting that the repression of telomerase activity is transcriptional. The tetomerase transcript has been shown to have at least six alternate splicing sites (four insertion sites and two deletion sites), and variants containing both or either of the deletion sites are present during development and in a panel of cancer cell lines we surveyed. One deletion (β site) and all four insertions cause premature translation terminations, whereas the other deletion (α site) is 36 bp and lies within reverse transcriptase (RT) motif A, suggesting that this deletion variant may be a candidate as a dominant-negative inhibitor of telomerase. We have cloned three alternately spliced hTERT variants that contain the α, β or both α and β deletion sites. These alternate splicing variants along with empty vector and wild-type hTERT were introduced into normal human fibroblasts and several telomerase-positive immortal and tumor cell lines. Expression of the α site deletion variant (hTERT α-) construct was confirmed by Western blotting. We found that none of the three alternate splicing variants reconstitutes telomerase activity in fibroblasts. However, hTERT α-inhibits telomerase activities in telomerase-positive cells, causes telomere shortening and eventually cell death. This alternately spliced dominant-negative variant may be important in understanding telomerase regulation during development, differentiation and in cancer progression. Neoplasia (2000) 2, 433-440.

Original languageEnglish (US)
Pages (from-to)433-440
Number of pages8
JournalNeoplasia
Volume2
Issue number5
StatePublished - 2000

Fingerprint

Telomerase
Alternative Splicing
Telomere Shortening
RNA-Directed DNA Polymerase
Fibroblasts
Neoplasms
Tumor Cell Line
Cell Death
Complementary DNA
Chromosomes
Western Blotting
Cell Line

Keywords

  • Alternate splicing
  • Apoptosis
  • Dominant-negative inhibitor
  • Human telomerase
  • Telomere

ASJC Scopus subject areas

  • Cancer Research

Cite this

Yi, X., White, D. M., Aisner, D. L., Baur, J. A., Wright, W. E., & Shay, J. W. (2000). An alternate splicing, variant of the human telomerase catalytic subunit inhibits telomerase activity. Neoplasia, 2(5), 433-440.

An alternate splicing, variant of the human telomerase catalytic subunit inhibits telomerase activity. / Yi, X.; White, D. M.; Aisner, D. L.; Baur, J. A.; Wright, W. E.; Shay, J. W.

In: Neoplasia, Vol. 2, No. 5, 2000, p. 433-440.

Research output: Contribution to journalArticle

Yi, X, White, DM, Aisner, DL, Baur, JA, Wright, WE & Shay, JW 2000, 'An alternate splicing, variant of the human telomerase catalytic subunit inhibits telomerase activity', Neoplasia, vol. 2, no. 5, pp. 433-440.
Yi, X. ; White, D. M. ; Aisner, D. L. ; Baur, J. A. ; Wright, W. E. ; Shay, J. W. / An alternate splicing, variant of the human telomerase catalytic subunit inhibits telomerase activity. In: Neoplasia. 2000 ; Vol. 2, No. 5. pp. 433-440.
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